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中国精品科技期刊2020
管桐,高丽莎,隋得志,等. 天麻素调控SIRT3改善BV2细胞衰老和炎症的作用[J]. 食品工业科技,2022,43(19):410−418. doi: 10.13386/j.issn1002-0306.2022020026.
引用本文: 管桐,高丽莎,隋得志,等. 天麻素调控SIRT3改善BV2细胞衰老和炎症的作用[J]. 食品工业科技,2022,43(19):410−418. doi: 10.13386/j.issn1002-0306.2022020026.
GUAN Tong, GAO Lisha, SUI Dezhi, et al. Ameliorative Effect of Gastrodin on Aging and Inflammation of BV2 Cells by Regulating SIRT3[J]. Science and Technology of Food Industry, 2022, 43(19): 410−418. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022020026.
Citation: GUAN Tong, GAO Lisha, SUI Dezhi, et al. Ameliorative Effect of Gastrodin on Aging and Inflammation of BV2 Cells by Regulating SIRT3[J]. Science and Technology of Food Industry, 2022, 43(19): 410−418. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022020026.

天麻素调控SIRT3改善BV2细胞衰老和炎症的作用

Ameliorative Effect of Gastrodin on Aging and Inflammation of BV2 Cells by Regulating SIRT3

  • 摘要: 目的:探讨天麻素对D-半乳糖诱导的BV2细胞衰老的保护作用及机制。方法:使用不同浓度(10、20、30和40 μg/mL)的D-半乳糖刺激BV2细胞24 h,建立细胞衰老模型,并用CCK-8法筛选出D-半乳糖的最佳造模浓度;实验分为4组:正常组、模型组、Silent mating type information regulation 2 homolog 3(SIRT3)抑制剂+天麻素组和天麻素组;用CCK-8法检测不同浓度(10、20、30、40和50 μmol/L)的天麻素对D-半乳糖刺激的BV2细胞活力的影响,并筛选出最佳的天麻素浓度;使用β-半乳糖苷酶(Senescence β-Galactosidase,SA-β-Gal)染色检测细胞衰老面积;使用生化法检测各组细胞活性氧(Reactive oxygen species,ROS)水平;使用Enzyme linked immunosorbent assay(ELISA)法检测各组细胞神经炎症因子IL-1β(Interleukin 1β,IL-1β)、IL-6(Interleukin 6,IL-6)和TNF-α(Tumor necrosis factor α,TNF-α)水平;使用免疫荧光法检测各组细胞SIRT3荧光强度;使用Western Blot法检测细胞SIRT3、P16和P21的蛋白表达水平。结果:30 μg/mL的D-半乳糖刺激BV2细胞活力极显著降低(P<0.01),引起BV2细胞中SA-β-Gal染色面积和衰老蛋白P16和P21表达极显著增加(P<0.01),细胞中ROS水平和炎症因子IL-1β、IL-6和TNF-α水平极显著增高(P<0.01),细胞内SIRT3蛋白表达极显著降低(P<0.01)。而30 μmol/L天麻素能极显著提高D-半乳糖刺激的BV2细胞活力(P<0.01),并且极显著降低细胞SA-β-Gal染色面积和衰老蛋白P16和P21表达水平(P<0.01),极显著降低ROS水平和神经炎症因子IL-1β、IL-6和TNF-α水平(P<0.01),极显著提高细胞内SIRT3蛋白荧光强度和表达水平(P<0.01)。结论:天麻素能够提高D-半乳糖刺激的BV2细胞活力,改善BV2细胞衰老染色和衰老蛋白表达,并降低ROS水平和减缓炎症反应,这可能与天麻素提高SIRT3蛋白表达有关。

     

    Abstract: Objective: In order to explore the protective effects and mechanism of gastrodin on BV2 cells treated with D-galactose. Methods: The BV2 cells were treated with D-galactose at different concentrations (10, 20, 30 and 40 μg/mL) for 24 h to establish a senescent cell model, and the optimum concentration of D-galactose was selected by CCK-8 method; The cells were divided into control group, model group, silent mating type information regulation 2 homolog 3 (SIRT3) inhibitor+gastrodin group and gastrodin group; and the optimum concentration of D-galactose was selected by CCK-8 method; The effects of different concentrations of gastrodin (10, 20, 30, 40 and 50 μg/mL) on the viability of BV2 cells treated with D-galactose were detected by CCK-8 method, and the best concentration of gastrodin was selected; The aging area of BV2 cells was detected by Senescence β-Galactosidase staining (SA-β-Gal); The level of reactive oxygen species (ROS) in BV2 cells was detected by biochemical method; The levels of neuroinflammatory factor interleukin 1β (IL-1β), interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA); The fluorescence intensity of SIRT3 was detected by immunofluorescence method; The protein expression levels of SIRT3, P16 and P21 were detected by Western blot. Results: Treatment with D-galactose at 30 μg/mL exerted a significant inhibitory effect on BV2 cell viability, resulting in SA-β-Gal staining area and the expression of aging proteins P16 and P21 increased (P<0.01), ROS level and inflammatory factor IL-1β, IL-6 and TNF-α significantly increased (P<0.01), and the expression of SIRT3 protein and fluorescence intensity decreased in BV2 cells (P<0.01). 30 μmol/L gastrodin significantly increased the of BV2 cells viability treated with D-galactose (P<0.01); Gastrodin reduced SA-β-Gal staining area and the expression levels of aging proteins P16 and P21 (P<0.01); Gastrodin significantly decreased the level of ROS and neuroinflammatory factor IL-1β, IL-6 and TNF-α; The expression level of SIRT3 protein in cells was significantly increased (P<0.01). Treatment with gastrodin increased the fluorescence intensity and protein levels of SIRT3. Conclusion: Gastrodin increased the viability of the BV2 cells treated with D-galactose, improved the SA-β-Gal staining area and aging protein P16 and P21 expression, reduced the ROS level and slowed down the inflammatory response, which may be related to its increasing effect on the expression of SIRT3 protein.

     

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